While the acquisition of invasiveness is a critical step in early stage breast carcinomas (DCIS), no established molecular markers reliably identify tumor progression. The metastasis gene osteopontin is subject to alternative splicing, which yields 3 messages, osteopontin-a, osteopontin-b and osteopontin-c. Osteopontin-c is selectively expressed in invasive, but not in noninvasive, breast tumor cell lines, and it effectively supports anchorage independence. We evaluated osteopontin-c as a biomarker. The RNA message for osteopontin-c was present in 16 of 20 breast cancers (80%), but was undetectable in 22 normal specimens obtained from reduction mammoplasty. In contrast, osteopontin-a RNA was expressed at various levels in all 20 breast cancers, 11 tumor-surrounding tissues and 21 normal samples. The splice variant osteopontin-b was present at barely detectable levels in 18 of 20 cancers and in 6 of 22 normal breasts. By immunohistochemistry, 66 of 69 normal breasts were negative, while 3 showed low level staining. Among the breast cancers, 43 of 56 cores (77%) stained positive for osteopontin-c. When correlated with tumor grade, the staining for osteopontin-c increased from grade 1 to grade 3. In a total of 178 breast specimens analyzed, osteopontin-c was present in 78% of cancers, 36% of surrounding tissues and 0% of normal tissues. Furthermore, osteopontin-c detects a higher fraction of breast cancers than estrogen receptor (ER), progesterone receptor or HER2. In conjunction, osteopontin-c, ER and HER2 reliably predict grade 2-3 breast cancer. Hence, osteopontin-c is a diagnostic and prognostic marker that may have value in a diagnostic panel together with conventional breast cancer markers. ' 2007 Wiley-Liss, Inc.

Results suggest that 4E-BP1 overexpression is strongly associated with prostate cancer, especially when combined with PTEN and mTOR expression data. Hierarchical clustering analysis utilizing PTEN, mTOR, and 4E-BP1 separated normal from cancer cell populations in most cases.

Loss of major histocompatibility complex class II (MHC II)

No abstract

Analyses of biomarkers in tissues represent a critical component in the development of therapeutics. In oncology drug development, entire signaling pathways may be profiled with a variety of high‐throughput techniques. However, in the field of histology, tissue‐based diagnostics (IHC and ISH) are generally dominated by single‐ or dual‐analyte tests. In addition, manual methods for slide processing promote inconsistency, are labor demanding and time‐consuming. We have developed automated platforms for conducting IHC and ISH in widely available formalin‐fixed paraffin‐embedded (FFPE) tissues. These instruments enable rapid evaluation of gene and protein biomarkers in a high‐throughput format. Further, advances in detection, including Quantum dots (Qdots®), and hyper‐spectral imaging, provide a growing set of tools that complement biomarker studies. We have applied these methods to monitor markers that may be therapeutic targets, prognostic of disease, or surrogates of drug response. This report demonstrates the utility of the platform in the simultaneous detection and quantitation of breast cancer markers including, HER2, ER, PR, and Ki67. In addition to automation, spectral imaging and deconvolution algorithms were critical to the reproducible determination of multiplexed protein expression levels in FFPE samples.